Fc-Binding Peptides for Antibody Purification

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Solution Overview

Problem

Current antibody-binding molecules lack sufficient molecular diversity and stability, particularly in detecting, purifying, and immobilizing antibodies, due to limitations in binding specificity, affinity, and adaptability to various conditions, and existing techniques for analyzing non-native antibody structures are inefficient.

Innovation Solution

Development of novel polypeptides with specific amino acid sequences that exhibit high binding activity to the Fc region of immunoglobulin G, including those that bind selectively to non-native structures, allowing for efficient detection, purification, and removal of antibodies, and providing enhanced stability under various conditions without requiring cyclization.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If protein A derived from Staphylococcus is used for recovering and purifying antibodies, then binding activity to antibodies is achieved, but stability and production cost are insufficient

Engineering Contradiction:
ImprovestabilityVSAvoidproduction cost
Core Design Contradiction:
ReliabilityVSEase of manufacture

Solution Approach 1:

The patent uses short peptide sequences (10-20 amino acids) instead of large protein A molecules. These small peptides are chemically synthesized rather than produced through complex biological systems, making them cheaper and more stable. The peptides can be easily produced by standard peptide synthesis methods without requiring bacterial culture and purification processes.

Inventive Principle:
Principle #27Cheap short-living objects (Disposable)

Solution Approach 2:

The patent identifies and utilizes specific binding motifs within the larger protein A structure. By segmenting the functional binding region into smaller peptide sequences (such as the YDPRTGTWRSSIAYGGG sequence), the invention creates minimal functional units that retain antibody binding capability while eliminating the need for producing entire protein A molecules.

Inventive Principle:
Principle #1Segmentation

2Adaptability or versatility

If existing antibody-binding peptides are used, then some binding activity is achieved, but molecular diversity is insufficient

Engineering Contradiction:
Improvemolecular diversityVSAvoidbinding specificity
Core Design Contradiction:
Adaptability or versatilityVSReliability

Solution Approach 1:

The patent systematically varies parameters of the peptide sequences including amino acid composition, sequence length (10-20 residues), and structural features (linear vs. cyclic). By changing these parameters while maintaining the core binding motif YDPRTGTWRSSIAYGGG, the invention generates a diverse family of peptides with different binding characteristics suitable for various applications.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent develops a universal binding motif that can function in multiple contexts - linear and cyclic forms, different lengths, various amino acid substitutions - all maintaining antibody binding capability. This universal motif can be adapted for different antibody types and application requirements, providing versatility across multiple scenarios.

Inventive Principle:
Principle #6Universality (Multi-functionality)

3Stability of the object's composition

If cyclic peptides with disulfide bonds are used to enhance stability, then binding affinity is improved, but complexity of structure increases

Engineering Contradiction:
Improvestructural stabilityVSAvoidstructural complexity
Core Design Contradiction:
Stability of the object's compositionVSDevice complexity

Solution Approach 1:

The patent employs simple linear peptide sequences that do not require complex cyclization or disulfide bond formation. These short linear peptides are chemically simpler to synthesize and handle, yet maintain sufficient stability for their intended applications through their optimized amino acid sequences and appropriate buffer conditions.

Inventive Principle:
Principle #27Cheap short-living objects (Disposable)

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

The novel polypeptides demonstrate high binding affinity and stability, enabling effective detection, purification, and removal of antibodies, including those with non-native structures, and can operate under reductive conditions, improving the efficiency of antibody handling processes.

Implementation Method 1

a polypeptide exhibiting binding activity to an Fc region of immunoglobulin G... wherein the polypeptide comprises an amino acid sequence represented by formula (1)

Methodology Applied
Scientific EffectProtein-protein binding interaction: Adsorption

Data Source

PatentEP2949750B1Antibody-binding peptide
Publication Date: 2019.11.13 NATIONAL INSTITUTE OF ADVANCED INDUSTRIAL SCIENCE & TECHNOLOGY
  • EP2949750B1 patent drawingFigure 1(A)~1(F)
  • EP2949750B1 patent drawingFigure 2
  • EP2949750B1 patent drawingFigure 3

AI summary

The invention relates to polypeptides consisting of amino acid sequences represented by the following formulas 1 to 4, which have binding activity to an Fc region of immunoglobulin G and can be favorably used in detecting, purifying, immobilizing or removing an antibody, immunoglobulin G or a protein containing an Fc region of immunoglobulin G, Y-D-P-x-T-G-T-W-R-S-x-[IL] (SEQ ID NO: 1) (1) R-[QRS]-x-x-[GS]-Y-D-P-R-T-G-T-W-R-S-S-I-A-Y-G-G (SEQ ID NO: 10) (2) G-V-V-R-Q-W-S-G-x-x-x-x-x-x-x-x-R-S-S-I-A-Y-G-G (SEQ ID NO: 20) (3) D-A-A-W-H-L-G-E-L-V-W-A-T-Y-Y-D-P-E-T-G-T-W-x-P-D-W-x-x-M (SEQ ID NO: 23) (4) where x represents any amino acid residue; and amino acid residues within the square brackets indicate that any one of the amino acid residues is selected.